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A novel peptide, selected from phage display library of random peptides, can efficiently target into human breast cancer cell

机译：从随机肽的噬菌体展示库中选择的新型肽可以有效地靶向人乳腺癌细胞

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摘要

To develop a targeting vector for breast cancer biotherapy, MDA-MB-231 cell, a human breast cancer cell line, was co-cultured with pC89 (9 aa) phage display library of random peptides. In multiple independent peptide-presenting phage screening trials, subtilisin was used as a protease to inactivate extra-cellular phages. The internalized phages were collected by cell lysising and amplified in E. coli XLI-Blue. Through five rounds of selection, the peptide-presenting phages which could be internalized in MDA-MB-231 cells were isolated. A comparison was made between internalization capacities of peptide-presenting phages isolated from MDA-MB-231 cells and RGD-integrin binding phage by coculturing them with other human tumor cell lines and normal cells. The nucleotide sequences of isolated peptide-presenting phages were then determined by DNA sequencing. To uncover whether phage coat protein or amino acid order was required for the character of the peptide to MDA-MB-231 cells, three peptides were synthesized. They are CASPSGALRSC, ASPSGALRS and CGVIFDHSVPC (the shifted sequence of CASPSGALRSC), and after coculturing them with different cell lines, their targeting capacities to MDA-MB-231 cells were detected. These data suggested that the internalization process was highly selective, and capable of capturing a specific peptide from parent peptide variants. Moreover, the targeting internalization event of peptides was an amino acid sequence dependent manner. The results demonstrated the feasibility of using phage display library of random peptides to develop new targeting system for intracellular delivery of macromolecules, and the peptide we obtained might be modified as a targeting vector for breast cancer gene therapy.
机译：为了开发用于乳腺癌生物疗法的靶向载体，将人乳腺癌细胞系MDA-MB-231细胞与随机肽的pC89（9 aa）噬菌体展示文库共培养。在多个独立的肽呈递噬菌体筛选试验中，枯草杆菌蛋白酶被用作蛋白酶来灭活细胞外噬菌体。通过细胞裂解收集内化的噬菌体，并在大肠杆菌XLI-Blue中扩增。通过五轮选择，分离了可以内在MDA-MB-231细胞中的肽呈递噬菌体。通过与其他人肿瘤细胞系和正常细胞共培养，从MDA-MB-231细胞分离的肽呈递噬菌体和RGD-整联蛋白结合噬菌体的内在化能力进行了比较。然后通过DNA测序确定分离的肽呈递噬菌体的核苷酸序列。为了揭示针对MDA-MB-231细胞的肽的特征是否需要噬菌体外壳蛋白或氨基酸顺序，合成了三种肽。它们是CASPSGALRSC，ASPSGALRS和CGVIFDHSVPC（CASPSGALRSC的移位序列），在将它们与不同的细胞系共培养后，检测到它们对MDA-MB-231细胞的靶向能力。这些数据表明内在化过程是高度选择性的，并且能够从亲本肽变体中捕获特定的肽。此外，肽的靶向内化事件是氨基酸序列依赖性方式。结果证明了使用随机肽的噬菌体展示文库开发用于大分子细胞内递送的新靶向系统的可行性，并且我们获得的肽可能被修饰为乳腺癌基因治疗的靶向载体。

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